title: DNA Sequence Fragment Containing C to A Mutation as a Convenient Mutation Standard for DHPLC Analysis.
abstract: OBJECTIVE
  Denaturing high performance liquid chromatography (DHPLC) is a high throughput approach for screening DNA sequence variations. To assess oven calibration, cartridge performance, buffer composition and stability, the WAVE Low and High Range Mutation Standards are employed to ensure reproducibility and accuracy of the chromatographic analysis. The purpose of this study was to provide a cost-effective homemade mutation standard for DHPLC analysis.
  
  
  METHODS
  DHPLC was performed to evaluate different elution temperatures of a 374 bp DNA fragment with C>A mutation at position of 59 to achieve a peak profile similar to the Low Mutation Standard. In order to verify the reproducibility of the homemade mutation standard using DHPLC, 15 different experiments were performed to compare the homemade mutation standard, the WAVE Low Range Mutation Standard with a positive DNA control sample.
  
  
  RESULTS
  We identified a comparable elution temperature and a peak profile with the WAVE Low Range Mutation Standard.
  
  
  CONCLUSIONS
  This study confirmed the reproducibility of the peak profile of our homemade mutation standard compared to the Low Mutation Standard using DHPLC analysis.
authors.affiliation: Department of Medical Genetics, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Hassan
authors.initials: H
authors.lastname: Dastsooz
authors.forename: Nazanin
authors.initials: N
authors.lastname: Vahedi
authors.forename: Majid
authors.initials: M
authors.lastname: Fardaei
keywords: ATP7B; DHPLC; Low range mutation stan- dard